Betulinic acid modulates the proliferation,invasion and collagen synthesis in keloid fibroblasts by regulating Wnt/β-catenin signaling pathway
Objective To investigate the effect of betulinic acid(BA)on the proliferation,invasion and collagen synthesis in keloid fibroblasts and explore the molecular mechanism.Methods The fibroblasts were isolated from keloid tissues using tissue explants adherent method.Cells were treated and divided into three groups including blank group,dimethyl sulfoxide(DMSO)group,and BA group.Cell growth was detected by cell counting kit-8(CCK-8)assay.Cell proliferation was evaluated by 5-ethynyl-2'-deoxyuridine(EdU).Cell apoptosis was measured by flow cytometry.Cell invasion was examined by Transwell assay.The activity of Wnt/β-catenin signaling pathway was monitored by luciferase reporter assay.The mRNA expression of c-myc and cyclin D1 was examined by reverse transcription quantitative real-time PCR(RT-qPCR).The protein expressions of Collagen type Ⅰ(Col Ⅰ),β-catenin,c-myc and cyclin D1 were detected by Western blot.Results Compared with blank group and DMSO group,BA-treated keloid fibroblasts had significantly decreased proliferation,increased the apoptosis,down-regulated the invasion,and reduced collagen synthesis(P<0.05).In addition,BA treated keloid fibroblasts had significantly reduced the activity of Wnt/β-catenin signaling pathway,and decreased the expression of β-catenin,c-myc and cyclin D1(P<0.05).Conclusion BA suppresses the proliferation,invasion and collagen synthesis in keloid fibroblasts through down-regulation of Wnt/β-catenin signaling pathway.
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